Harvesting murine alveolar macrophages and evaluating cellular activation induced by polyanhydride nanoparticles

Ana V. Chavez-Santoscoy, Lucas M. Huntimer, Amanda E. Ramer-Tait, Michael Wannemuehler, Balaji Narasimhan

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

Biodegradable nanoparticles have emerged as a versatile platform for the design and implementation of new intranasal vaccines against respiratory infectious diseases. Specifically, polyanhydride nanoparticles composed of the aliphatic sebacic acid (SA), the aromatic 1,6-bis(p-carboxyphenoxy)hexane (CPH), or the amphiphilic 1,8-bis(p-carboxyphenoxy)-3,6-dioxaoctane (CPTEG) display unique bulk and surface erosion kinetics1,2 and can be exploited to slowly release functional biomolecules (e.g., protein antigens, immunoglobulins, etc.) in vivo3,4,5. These nanoparticles also possess intrinsic adjuvant activity, making them an excellent choice for a vaccine delivery platform6,7,8. In order to elucidate the mechanisms governing the activation of innate immunity following intranasal mucosal vaccination, one must evaluate the molecular and cellular responses of the antigen presenting cells (APCs) responsible for initiating immune responses. Dendritic cells are the principal APCs found in conducting airways, while alveolar macrophages (AM) predominate in the lung parenchyma9,10,11. AM are highly efficient in clearing the lungs of microbial pathogens and cell debris12,13. In addition, this cell type plays a valuable role in the transport of microbial antigens to the draining lymph nodes, which is an important first step in the initiation of an adaptive immune response9. AM also express elevated levels of innate pattern recognition and scavenger receptors, secrete pro-inflammatory mediators, and prime naïve T cells12,14. A relatively pure population of AM (e.g., greater than 80%) can easily be obtained via lung lavage for study in the laboratory. Resident AM harvested from immune competent animals provide a representative phenotype of the macrophages that will encounter the particle-based vaccine in vivo. Herein, we describe the protocols used to harvest and culture AM from mice and examine the activation phenotype of the macrophages following treatment with polyanhydride nanoparticles in vitro.

Original languageEnglish (US)
Article numberA47
JournalJournal of Visualized Experiments
Issue number64
DOIs
StatePublished - Jun 8 2012

Keywords

  • AM
  • Activation
  • Alveolar macrophages
  • Bioengineering
  • Harvesting
  • Issue 64
  • Lung lavage
  • Microbiology
  • Nanoparticles
  • Polyanhydride

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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